Apparatus for filtration-leukopheresis for separation and concentration of human granulocytes

ABSTRACT

Apparatus of simple character is provided for continuous withdrawal of blood from a human donor, forced extracorporeal circulation of blood of the donor with separation of granulocytes, and return by gravity of the leukocyte-poor whole blood to the donor.

United States Patent Djerassi *July 1, 1975 APPARATUS FOR 3,014,48ll2/l96l Rumble et al. 128/214 F FILTRATION LEUKOPHERESIS FOR glarkk. 1.

, wan SEPARATION AND CONCENTRATION OF 3,462,361 8/1969 Greenwalt et al.210/23 HUMAN GRANULOCYTES 3,648,694 3 1972 Mogos et a1 1. 128/214 F [76]Inventor: Isaac Djerassi, 2034 Delancey FL,

Philadelphia, Pa. 19103 FOREIGN PATENTS OR APPLICATIONS 1 Notice: h p iof the term of this Germany l28/DIG. I2

patent subsequent to Apr. 9, 1991, has been d'sclalmed' PrimaryExaminerDalton L. Truluck [22] Fil d; A 2, 1973 Attorney, Agent, orFirm-Zachary T. Wobensmith,

2 d;Z h T.W 'th,]] 21 Appl. No.: 346,729 ac wens I [52] US. Cl. 128/214R; 128/276 [57] ABSTRACT [5 1) Int. Cl Afilm 01/03 [58] Field Search128/2131 214 F1 Apparatus ofsimple character is provided for continu-28/214 B1 214 C1 2l4-21 278; ous withdrawal of blood from a human donor,forced 23/2585? 210/23 extracorporeal circulation of blood of the donorwith separation of granulocytes, and return by gravity of [56]References cued the leukocyte-poor whole blood to the donor.

UNITED STATES PATENTS 2,876,769 3/1959 Cordova 23/2585 8 Claims, 2Drawing Figures n lI APPARATUS FOR FlLTRATION-LEUKOPIIERESIS FORSEPARATION AND CONCENTRATION OF HUMAN GRANULOCYTES BACKGROUND OF THEINVENTION 1. Field of the Invention This invention relates to apparatusfor filtrationleukopheresis for separation and concentration of largeamounts of normal human granulocytes.

2. Description of the Prior Art It has heretofore been proposed toextract blood from a human donor for use as whole blood, for separationof plasma, for separation of granulocytes or other blood componentsprimarily for the purpose of transfusion to individuals in need of suchcomponents.

Blood removal and collection apparatus has taken various forms andreference may be had to the U.S. Pat. Nos. to Strumia, 2,845,929,Gewecke et al., 2,757,669, Welch, Jr., 2,982,286, Rieutord et al.,2,757,375, Poitras, 2,784,932 Erikson, 2,597,715, and Judson et al.,3,489,145.

The delivery of blood plasma and the like to a patient for transfusioncan be effected by gravity or by applied pressure as shown in the U.S.Pat. No. to Rundhaug, 2,842,123.

The harvesting of normal granulocytes with special continuous flowcentrifugation (Freirach, E. J., et al., Separation and collection ofleukocytes. Cancer Res. 1516-1520 1965) is inefficient and impracticalfor widespread use.

The batch treating of whole blood to remove leukocytes with a filter isshown in the US. Pat. No. to Greenwalt, 3,462,361.

It has heretofore been undertaken to withdraw blood from a donor, in aplastic bag with saline solution and a mixture of heparin, saline andsodium citrate selectively delivered to the plastic bag and thentransferring the fluid through two leukocyte filters and then advancethe blood by manual manipulation to another plastic bag for returnthrough a tube to the donor, a second source of saline solution beingconnected to the return tube. The respective tubes were controlled byhemostats for manual regulation and the entire procedure was manual.

Very close supervision and control by trained personnel has beenrequired and the time for the filtrationleukopheresis has required aboutsome 4 hours or more with each individual donor.

In my prior application for US. Letters Pat. filed May 18, 1972, Ser.No. 254,609, now US. Pat. No. 3,802,432 issued Apr. 9, 1974, there isdescribed apparatus for separation and retention of human granulocyteswith which the weight of the blood in intermediate receptacles in itscircuit from and to the body of the donor is utilized to control theflow.

In my prior application for US. Letters Pat. filed Dec. 20, 1972, Ser.No. 317,057, improved apparatus is shown.

The present invention comprises simplified apparatus utilizing in oneembodiment constant volume of blood from the donor and to and throughthe granulocyte separating filter and in another embodiment constantpressure of the blood from the donor to and through the filter in bothinstances for effecting the flow through the apparatus to a receptaclefor gravity return of leukocyte-poor whole blood to the donor.

SUMMARY OF THE INVENTION In accordance with the present inventionapparatus is provided for harvesting granulocytes from a single donorand within a relatively short period of time and which includes a filterconnected to a vented blood return receptacle, the blood being deliveredto and through the filter to the return receptacle either by constantvolume or by constant pressure delivery to the filter.

It is the principal object of the invention to provide apparatus forharvesting granulocytes from human blood of a donor with return of theleukocyte-poor blood to the donor which is of simplified construction,which requires a minimum of attention and which will permit of rapidcycling and attendant short period of retention of the donor.

It is a further object of the invention to provide apparatus of thecharacter aforesaid in which the filter and the blood return receptacleand associated tubing may be supplied as a disposable unit forconnection to a permanently installed withdrawal and delivery apparatus.

It is a further object of the invention to provide apparatus of thecharacter aforesaid in which the withdrawal and delivery apparatus mayselectively be such as to effect a constant volume or a constantpressure transfer of the blood from the donor.

Other objects and advantageous features of the invention will beapparent from the description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS The nature and characteristic featuresof the inven tion will be more readily understood from the followingdescription taken in connection with the accompanying drawings formingpart hereof, in which:

FIG. 1 is a diagrammatic view of a preferred form of apparatus inaccordance with the invention utilizing constant pressure for blooddelivery; and

FIG. 2 is a diagrammatic view of another preferred form of apparatus inaccordance with the invention, utlizing constant volume for blooddelivery.

It should, of course, be understood that the description and drawingsherein are illustrative merely and that various modifications andchanges can be made in the structure disclosed without departing fromthe spirit of the invention.

Like numerals refer to like parts throughout the several views.

DESCRIPTION OF THE PREFERRED EMBODIMENT Referring now more particularlyto FIG. I of the drawings, a fluid connection 10 of flexible tubing, hasa needle (not shown) of well known type, and preferably of 15 gauge, onits free end for insertion into a vein in one arm A1 of the donor forwithdrawing blood.

The fluid connection 19 preferably extends to branch connections 11 and12 of flexible tubing and past inlet control valves 13 and I4, which maybe non-return valves or positively positioned valves. The valves 13 and14 preferably act externally of and are adapted to pinch or release thetubes 11 and 12. The pipes 11 and 12 extend to flexible blood receivingreceptacles I5 and 16, preferably of inert transparent or translucentsynthetic plastic, suspended from fixed brackets 17 and 18 in rigidenclosing cylinders 19 and 20. The cylinders l9 and 20 are preferablytransparent for observation of the receptacles l and 16 and each has itsinterior alternately placed under vacuum and under pressure and phasedso that when cylinder 19 is under vacuum cylinder 20 is under pressure.

Vacuum pipes 23 and 24, connected to a suitable source of vacuum VP andconnected to the cylinders 19 and 2.0 through control valves 25 and 26and pressure supply pipes 27 and 28, connected to a suitable source ofpressure PP through control valves 29 and 30, permit alternate pressureand vacuum application in cylinders l9 and 20.

The receptacles l5 and 16 are connected by fluid delivery connections 33and 34, of flexible tubing, past control valves 35 and 36, which may benon-return valves or positively positioned valves. The valves 35 and 36preferably act externally of and are adapted to pinch or release thetubes 33 and 34. The pipes 33 and 34 extend to a pipe 37 which isconnected to a filter 38 which delivers blood either directly to pipe 44or through pipe 39 to and through a transparent chamber 40 in which theflow of blood can be observed directly. The filter 38 can be of anysuitable type which separates and retains granulocytes and permits thepassage of leukocyte-poor whole blood for return to the donor. Filtersidentified as LeukoPak Leukocyte Filter available from FenwalLaboratories, Division of Travenol Laboratories, lnc., Morton Grove,Illinois, and which utilize nylon fibers for adherence thereto of thegranulocytes, have been found satisfactory.

The unit comprising the pendant filter 38 and chamber 40, if used, arepreferably supported by a stand 41 having a hook 42 for detachableengagement by a ring 43 on the bag 37.

The filter assembly 38 and 40 is connected by a pipe 44 to the lowerpart of a vented blood return receptacle 45.

The receptacle 45 is preferably supported on the stand 41 by a hook 47for detachable engagement by a ring 48 on the receptacle 45.

The receptacle 45 has an air vent tube 49 extending upwardly to close tothe top of the receptacle 45 to prevent pressure build-up within thereceptacle 45.

The receptable 45 is connected through a transparent sight tube 53 by aflexible tube 52 which has a needle (not shown) of well known type andpreferably of 15 gauge on its free end for insertion into a vein in theother arm A2 of the donor for return, by gravity of granulocyte-poorwhole blood to the donor.

The structure, irreversibly connected, comprising the fluid connection10, tubes 11 and 12, receptacles l5 and I6, pipes 33 and 34, pipe 37,filter 38 with or without chamber 40, pipe 44, vented receptacle 45 andpipe 52 with its sight tube 53 can be made as a single use unit one foreach donor pre-sterilized and in a sealed protective package. Thenecessity for breaking sterile points of connection at the time of usagewill thus be avoided. Quick removal and replacement can be effected.

Referring now to FIG. 2 of the drawings, a fluid connection 10 offlexible tubing is provided, as before, to a needle for withdrawingblood from a vein in one arm A1 of the donor.

The fluid connection 10 is connected to the inlet side of a variablespeed positive displacement liquid pump 60. The pump 60, as a positivedisplacement pump, will have a constant volume delivery per revolutionirrespective of the speed of rotation and can have a revolution counter61 connected thereto and calibrated, if desired, in accordance with theliquid volume delivery of the pump 60.

The delivery side of the pump is connected by a flexible pipe 62 to afilter 38, of the type previously described, for delivery to a ventedblood return receptacle 45.

The receptacle 45 is connected by a flexible tube 52 for return bygravity, of granulocyte-poor whole blood to a needle in a vein in thearm A2 of the donor.

When suction of the blood through filter 38 by the pump is preferred,the pump is located to apply its action on pipe 44. In this case pipe 44may be of special material with increased resiliency such as siliconizedrubber or plastic.

The structure, irreversibly connected, comprising the pipe 62, filter38, with or without chamber 40, pipe 44, vented receptacle 45, and pipe52 with or without a transparent chamber, can be made as a single useunit similar to that previously described.

In the form of the invention shown in FlG. l, the blood is withdrawnfrom a vein in the arm A1 of the donor by the suction alternatelycreated in the cylinders l9 and 20 and therein on the flexiblereceptacles l5 and 16 and therefrom in the fluid connection 10. Theblood in the receptacles l5 and 16 is then alternately delivered underpressure through the pipe 37, through the filter 38 under pressure andthrough the pipe 39 and chamber 40 under pressure which pressure byreason of the alternating delivery from the receptacles l5 and 16 issubstantially constant.

In the filter 38 the granulocytes are separated from the whole blood,and collected in the filter 38 for subsequent recovery, theleukocyte-poor whole blood still under pressure being delivered to thevented receptacle 45.

The leukocyte-poor blood is then returned by gravity to a vein in thearm A2 of the donor through the return pipe 52, the height of thereceptacle 45 being such as to insure adequate rate of return of theblood and the receptacle 45 providing the desired storage capacity forcontinuous return of blood.

In the form of the invention shown in FIG. 2, the blood is withdrawnfrom a vein in the arm A1 of the donor by the suction of the pump 60 andcontinuously delivered with constant volume through the pipe 62 to thefilter 38.

The blood is delivered to and through the filter 38 under pressure or bysuction through the filter 38 by the continuity of flow and to thevented receptacle 45, as before.

The granulocytepoor whole blood in the receptacle 45 is returned bygravity to a vein of the donor as previously explained.

The quantity of blood delivered by the pump 60, can be measured by thecounter 61 which functions as a volume meter.

I claim:

1. Apparatus for filtration-leukopheresis comprising a venous bloodsupply connection adapted to be connected to a donor,

a venous blood return connection adapted to be connected to the donor,

a leukocyte separating and retaining filter interposed in series withsaid connections and in fluid communication therewith for extractingleukocytes from blood passing therethrough.

a permanently vented receptacle in continuous communication with theatmosphere interposed in series between said filter and said returnconnection and in fluid communication therewith.

fixedly mounted detachable supporting members for said filter and saidvented receptacle. and

power driven pump means upstream of said receptacle for continuouslydelivering blood from said supply connection to and through said filterand to said vented receptacle.

2. The combination defined in claim 1 in which said last mentioned meansincludes members for impelling said blood under constant positive airpressure.

3. The combination defined in claim I in which said last mentioned meansincludes a plurality of flexible receptacles for alternate receipt anddelivery of blood from the donor. and pressure and vacuum applying meansfor activating said flexible receptacles. 4. The combination defined inclaim 1 in which said last mentioned means includes a constant volumecontinuous impelling menber. 5. The combination defined in claim 4 inwhich said impelling member is a constant volume pump. 6. Thecombination defined in claim 5 in which said pump has a counter actuatedthereby. 7. The combination defined in claim 1 in which said filter andsaid receptacle are interconnected and have supply and deliveryconnections to provide a replaceable unit for individual use by a donor.8. A replaceable unit as defined in claim 7 in which said supply pipehas a plurality of flexible blood receiving receptacles interposedtherein.

1. Apparatus for filtration-leukopheresis comprising a venous bloodsupply connection adapted to be connected to a donor, a venous bloodreturn connection adapted to be connected to the donor, a leukocyteseparating and retaining filter interposed in series with saidconnections and in fluid communication therewith for extractingleukocytes from blood passing therethrough, a permanently ventedreceptacle in continuous communication with the atmosphere interposed inseries between said filter and said return connection and in fluidcommunication therewith, fixedly mounted detachable supporting membersfor said filter and said vented receptacle, and power driven pump meansupstream of said receptacle for continuously delivering blood from saidsupply connection to and through said filter and to said ventedreceptacle.
 2. The combination defined in claim 1 in which said lastmentioned means includes members for impelling said blood under constantpositive air pressure.
 3. The combination defined in claim 1 in whichsaid last mentioned means includes a plurality of flexible receptaclesfor alternate receipt and delivery of blood from the donor, and pressureand vacuum applying means for activating said flexible receptacles. 4.The combination defined in claim 1 in which said last mentioned meansincludes a constant volume continuous impelling menber.
 5. Thecombination defined in claim 4 in which said impelling member is aconstant volume pump.
 6. The combination defined in claim 5 in whichsaid pump has a counter actuated thereby.
 7. The combination defined inclaim 1 in which said filter and said receptacle are interconnected andhave supply and delivery connections to provide a replaceable unit forindividual use by a donor.
 8. A replaceable unit as defined in claim 7in which said supply pipe has a plurality of flexible blood receivingreceptacles interposed therein.